【Abstract】ObjectiveSome studies have demonstrated that recombinant adenoviruses are efficient vectors for gene transfer to the venous wall and AdCMV.tk encoded thymidine kinase can be used to reduce restenosis. In this study AdCMV.tk was apply to human vein smooth muscle cells (SMC) and organ cultured saphenous veins to study its effects on proliferation of SMCs and reduction of intimal hyperplasia.
MethodsThe adenovirus vector transferred tk gene and mark gene lacZ to the SMC of human saphenous veins and organ cultured vein segments. Various concentrations ganciclovir (GCV) were contained in culture media. The efficiency of gene transfer was studied by using Xgal staining. The proliferation of SMC was monitored by the method of trypan blue exclusion. The bystander effect was observed by mixed cell culture. After vein segments treated by AdCMV.tk+GCV and cultured for 14 days, HE and VG staining were carried out and intimal thickness was analysis by computer image system.
ResultsAdenovirus vector could infect saphenous vein SMC efficiently both in cultured SMCs and organ cultured vein segments. Gene expression sustained 14 d at least. The inhibition of SMCs proliferation in vitro was a positive correlation in GCV concentrations and the levels of tk expression. The proliferation of SMCs transfectered lacZ wasn’t restrained by GCV (P＜0.05). In mixed cell experiment there was at least 55% reduction in total cell number when as few as 10% of the cells express tk. Assessment of this “suicide gene strategy” in saphenous vein organ culture model demonstrated that veins treated with AdCMV.tk+GCV had a significant reduction at 14 days in the intimal thickness compared to control group (P＜0.01).
ConclusionThe results suggest that adenovirusmediated gene transfer of tk along with GCV administration may be a useful strategy to treat the proliferation of intimal hyperplasia of transplanting saphenous veins. Bystander effects are amplified by AdCMV.tk/GCV gene therapy system.
Citation：
LUO Tao,ZHANG Qiang,ZHANG Jian,LI Jianxin,GU Yongquan,YU Hengxi,WANG Zhonggao.. The Inhibitive Effects of Adenovirus Mediated tk Gene Transfer on Smooth Muscle Cells Proliferation and Intimal Hyperplasia. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2006, 13(3): 291-294. doi:
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- 1. Davies MG, Hagen PO. Pathophysiology of vein graft failure: a review [Ｊ]. Eur J Vasc Endovasc Surg， 1995; 9(1)∶7.
- 2. Ip JH, Fuster V, Badimon L, et al. Syndromes of accelerated atherosclerosis: role of vascular injury and smooth muscle cell proliferation [Ｊ]. J Am Coll Cardiol， 1990; 15(7)∶1667.
- 3. Bulkley BH, Hutchins GM. Accelerated “atherosclerosis”. A morphologic study of 97 saphenous vein coronary artery bypass grafts [Ｊ]. Circulation， 1977; 55(1)∶163.
- 4. Guzman RJ, Hirschowitz EA, Brody SL, et al. In vivo suppression of injuryinduced vascular smooth muscle cell accumulation using adenovirusmediated transfer of the herpes simplex virus thymidine kinase gene [Ｊ]. Proc Natl Acad Sci USA， 1994; 91(22)∶10732.
- 5. 罗涛， 张强， 刁彦鹏， 等. 腺病毒载体转染人大隐静脉平滑肌细胞效率的研究 [Ｊ]. 中华实验外科杂志, 2003； 20（3）∶209.
- 6. Francis SC, Raizada MK, Mangi AA, et al. Genetic targeting for cardiovascular therapeutics: are we near the summit or just beginning the climb? [Ｊ]. Physiol Genomics, 2001; 7(2)∶79.
- 7. van Dillen IJ, Mulder NH, Vaalburg W, et al. Influence of the bystander effect on HSVtk/GCV gene therapy. A review [Ｊ]. Curr Gene Ther, 2002; 2(3)∶307.
- 8. Channon KM, Fulton GJ, Gray JL, et al. Efficient adenoviral gene transfer to early venous bypass grafts: comparison with native vessels [Ｊ]. Cardiovasc Res， 1997; 35(3)∶505.
- 9. Borrelli E, Heyman R, Hsi M, et al. Targeting of an inducible toxic phenotype in animal cells [Ｊ]. Proc Natl Acad Sci USA， 1988; 85(20)∶7572.
- 10. Robe PA, Jolois O, N’Guyen M, et al. Modulation of the HSVTK/ganciclovir bystander effect by nbutyrate in glioblastoma: correlation with gapjunction intercellular communication [Ｊ]. Int J Oncol， 2004; 25(1)∶187.
- 11. Soyombo AA, Angelini GD, Bryan AJ, et al. Intimal proliferation in an organ culture of human saphenous vein [Ｊ]. Am J Pathol， 1990; 137(6)∶1401.
- 12. Ojha M. Wall shear stress temporal gradient and anastomotic intimal hyperplasia [Ｊ]. Circ Res， 1994; 74(6)∶1227.
